Multifunctional nanoagent for thrombus-targeted fibrinolytic therapy

Jason R. McCarthy, Irina Y. Sazonova, S. Sibel Erdem, Tetsuya Hara, Brian D. Thompson, Purvish Patel, Ion Botnaru, Charles P. Lin, Guy L. Reed, Ralph Weissleder, Farouc A. Jaffer

Research output: Contribution to journalArticle

39 Citations (Scopus)

Abstract

Background: Current thrombolytic therapies utilize exogenous plasminogen activators (PAs) to effectively lyse clots, restoring blood flow, and preventing tissue and organ death. These PAs may also impair normal hemostasis, leading to life-threatening bleeding, including intracerebral hemorrhage. Aims: This study aims to develop new thrombus-targeted fibrinolytic agents that harness the multifunctional theranostic capabilities of nanomaterials, potentially allowing for the generation of efficacious thrombolytics while minimizing deleterious side effects. Materials & methods: A thrombus-targeted nano-fibrinolytic agent was synthesized using a magnetofluorescent crosslinked dextran-coated iron oxide nanoparticle platform that was conjugated to recombinant tissue PA (tPA). Thrombus-targeting was achieved by derivatizing the nanoparticle with an activated factor XIII (FXIIIa)-sensitive peptide. Human plasma clot binding ability of the targeted and control agents was assessed by fluorescence reflectance imaging. Next, the in vitro enzymatic activity of the agents was assessed by S2288-based amidolytic activity, and an ELISA D-dimer assay for fibrinolysis. In vivo targeting of the nanoagent was next examined by intravital fluorescence microscopy of murine arterial and venous thrombosis. The fibrinolytic activity of the targeted nanoagent compared to free tPA was then evaluated in vivo in murine pulmonary embolism. Results: In vitro, the targeted thrombolytic nanoagent demonstrated superior binding to fresh-frozen plasma clots compared to control nanoagents (analysis of variance, p < 0.05). When normalized by S2288-based amidolytic activity, targeted, control and free tPA samples demonstrated equivalent in vitro fibrinolytic activity against human plasma clots, as determined by ELISA D-dimer assays. The FXIIIa targeted fibrinolytic nanoagent efficiently bound the margin of intravascular thrombi as detected by intravital fluorescence microscopy. In in vivo fibrinolysis studies the FXIIIa-targeted agent lysed pulmonary emboli with similar efficacy as free tPA (p > 0.05). Conclusion: The applicability of a FXIIIa-targeted thrombolytic nanoagent in the treatment of thromboembolism was demonstrated in vitro and in vivo. Future studies are planned to investigate the safety profile and overall efficacy of this class of nanoagents. Original submitted: 8 July 2011; Revised submitted: 16 November 2011; Published online 21 February 201.

Original languageEnglish (US)
Pages (from-to)1017-1028
Number of pages12
JournalNanomedicine
Volume7
Issue number7
DOIs
StatePublished - Jul 1 2012

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Fibrinolytic Agents
Plasminogen Activators
Thrombolytic Therapy
Thrombosis
Factor XIIIa
Tissue
Nanoparticles
Plasma (human)
Fluorescence microscopy
Tissue Plasminogen Activator
Analysis of variance (ANOVA)
Dextrans
Nanostructured materials
isoleucyl-prolyl-arginine-4-nitroanilide
Assays
Blood
Fluorescence
Plasmas
Imaging techniques
Peptides

All Science Journal Classification (ASJC) codes

  • Bioengineering
  • Medicine (miscellaneous)
  • Biomedical Engineering
  • Materials Science(all)

Cite this

McCarthy, J. R., Sazonova, I. Y., Erdem, S. S., Hara, T., Thompson, B. D., Patel, P., ... Jaffer, F. A. (2012). Multifunctional nanoagent for thrombus-targeted fibrinolytic therapy. Nanomedicine, 7(7), 1017-1028. https://doi.org/10.2217/nnm.11.179

Multifunctional nanoagent for thrombus-targeted fibrinolytic therapy. / McCarthy, Jason R.; Sazonova, Irina Y.; Erdem, S. Sibel; Hara, Tetsuya; Thompson, Brian D.; Patel, Purvish; Botnaru, Ion; Lin, Charles P.; Reed, Guy L.; Weissleder, Ralph; Jaffer, Farouc A.

In: Nanomedicine, Vol. 7, No. 7, 01.07.2012, p. 1017-1028.

Research output: Contribution to journalArticle

McCarthy, JR, Sazonova, IY, Erdem, SS, Hara, T, Thompson, BD, Patel, P, Botnaru, I, Lin, CP, Reed, GL, Weissleder, R & Jaffer, FA 2012, 'Multifunctional nanoagent for thrombus-targeted fibrinolytic therapy', Nanomedicine, vol. 7, no. 7, pp. 1017-1028. https://doi.org/10.2217/nnm.11.179
McCarthy JR, Sazonova IY, Erdem SS, Hara T, Thompson BD, Patel P et al. Multifunctional nanoagent for thrombus-targeted fibrinolytic therapy. Nanomedicine. 2012 Jul 1;7(7):1017-1028. https://doi.org/10.2217/nnm.11.179
McCarthy, Jason R. ; Sazonova, Irina Y. ; Erdem, S. Sibel ; Hara, Tetsuya ; Thompson, Brian D. ; Patel, Purvish ; Botnaru, Ion ; Lin, Charles P. ; Reed, Guy L. ; Weissleder, Ralph ; Jaffer, Farouc A. / Multifunctional nanoagent for thrombus-targeted fibrinolytic therapy. In: Nanomedicine. 2012 ; Vol. 7, No. 7. pp. 1017-1028.
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N2 - Background: Current thrombolytic therapies utilize exogenous plasminogen activators (PAs) to effectively lyse clots, restoring blood flow, and preventing tissue and organ death. These PAs may also impair normal hemostasis, leading to life-threatening bleeding, including intracerebral hemorrhage. Aims: This study aims to develop new thrombus-targeted fibrinolytic agents that harness the multifunctional theranostic capabilities of nanomaterials, potentially allowing for the generation of efficacious thrombolytics while minimizing deleterious side effects. Materials & methods: A thrombus-targeted nano-fibrinolytic agent was synthesized using a magnetofluorescent crosslinked dextran-coated iron oxide nanoparticle platform that was conjugated to recombinant tissue PA (tPA). Thrombus-targeting was achieved by derivatizing the nanoparticle with an activated factor XIII (FXIIIa)-sensitive peptide. Human plasma clot binding ability of the targeted and control agents was assessed by fluorescence reflectance imaging. Next, the in vitro enzymatic activity of the agents was assessed by S2288-based amidolytic activity, and an ELISA D-dimer assay for fibrinolysis. In vivo targeting of the nanoagent was next examined by intravital fluorescence microscopy of murine arterial and venous thrombosis. The fibrinolytic activity of the targeted nanoagent compared to free tPA was then evaluated in vivo in murine pulmonary embolism. Results: In vitro, the targeted thrombolytic nanoagent demonstrated superior binding to fresh-frozen plasma clots compared to control nanoagents (analysis of variance, p < 0.05). When normalized by S2288-based amidolytic activity, targeted, control and free tPA samples demonstrated equivalent in vitro fibrinolytic activity against human plasma clots, as determined by ELISA D-dimer assays. The FXIIIa targeted fibrinolytic nanoagent efficiently bound the margin of intravascular thrombi as detected by intravital fluorescence microscopy. In in vivo fibrinolysis studies the FXIIIa-targeted agent lysed pulmonary emboli with similar efficacy as free tPA (p > 0.05). Conclusion: The applicability of a FXIIIa-targeted thrombolytic nanoagent in the treatment of thromboembolism was demonstrated in vitro and in vivo. Future studies are planned to investigate the safety profile and overall efficacy of this class of nanoagents. Original submitted: 8 July 2011; Revised submitted: 16 November 2011; Published online 21 February 201.

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